高效液相色谱-串联质谱法测定金钗石斛中石斛碱和毛兰素的含量  

Determination of Dendrobine and Erianin in Dendrobium Nobile Lindl. by High Performance Liquid Chromatography-Tandem Mass Spectrometry

作　　者：吕谦 刘超 LÜQian;LIU Chao(Luzhou Market Inspection Testing Center,Luzhou 646000,China;Leshan Food and Drug Inspection Center,Leshan 614000,China)

机构地区：[1]泸州市市场检验检测中心,泸州646000 [2]乐山市食品药品检验检测中心,乐山614000

出　　处：《理化检验（化学分册）》2024年第11期1167-1172,共6页Physical Testing and Chemical Analysis(Part B:Chemical Analysis)

基　　金：乐山市重点研究项目(22ZDYJ0108)。

摘　　要：取粉碎均匀的石斛样品1 g置于50 m L聚四氟乙烯离心管中,加入0.1 mol·L^(-1)盐酸溶液5 m L,涡旋振荡2 min,充分浸润,加入15 m L乙醇,涡旋振荡1 min,超声30 min,冷却至室温,加入6 g无水硫酸镁和1.5 g无水乙酸钠,振摇3 min,冰浴冷却5 min,以转速8000 r·min^(-1)离心5 min,取上清液1 m L用乙醇定容至25 m L,采用高效液相色谱-串联质谱法(HPLC-MS/MS)测定石斛碱和毛兰素的含量。以Kinetex F5色谱柱为固定相,以不同体积比的乙醇-0.1%(体积分数)甲酸溶液混合液为流动相进行梯度洗脱,质谱分析采用电喷雾离子(ESI)源,正离子(ESI+)扫描模式,多反应监测(MRM)模式。结果表明,石斛碱和毛兰素的质量浓度在一定范围内与对应的峰面积呈线性关系,检出限(3S/N)分别为0.02,0.003 mg·kg^(-1);按照标准加入法进行回收试验,回收率为93.6%~97.1%,测定值的相对标准偏差(n=6)均小于3.0%;方法用于测定不同栽培方式、不同年限、不同部位金钗石斛中石斛碱、毛兰素的含量,均存在差异。The evenly crushed Dendrobium nobile Lindl.Sample(1 g)was taken,and placed in a 50 mLpolytetrafluoroethylene centrifuge tube.5 mL of 0.1 mol·L^(−1) hydrochloric acid solution was added,and the mixture was vortexed for 2 min to immerse fully.15 mL of ethanol was added,and the mixture was vortexed for 1 min,and sonicated for 30 min.After cooling to room temperature,6 g of anhydrous magnesium sulfate and 1.5 g of anhydrous sodium acetate were added.The mixture was shaken for 3 min,cooled in an ice bath for 5 min,and centrifuged at a rotational speed of 8000 r·min^(−1) for 5 min.1 mL of supernatant was taken and diluted to 25 mL with ethanol,and dendrobine and erianin were determined by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS).Kinetex F5 chromatographic column was used as the stationary phase and the mixed solution composed of ethanol and 0.1%(volume fraction)formic acid solution at different volume ratios was used as mobile phase for gradient elution.Electrospray ion(ESI)source was used for mass spectrometry.The targets were analyzed by multiple reaction monitoring(MRM)mode in positive(ESI+)scanning mode.As shown by the results,linear relationships between the corresponding peak areas and mass concentrations of dendrobine and erianin were found in definite ranges,with detection limits(3S/N)of 0.02,0.003 mg·kg^(−1),respectively.Test for recovery was made by the standard addition method,giving results in the range of 93.6%-97.1%,with RSDs(n=6)of the determined values less than 3.0%.This method was used to determine the dendrobine and erianin in different cultivation methods,different years,and different parts of Dendrobium nobile Lindl.,and there were differences among them.

关 键 词：高效液相色谱-串联质谱法(HPLC-MS/MS) 金钗石斛 石斛碱 毛兰素 

分 类 号：O657.63[理学—分析化学]